Protective Roles of Shilajit in Modulating Resistin, Adiponectin, and Cytokines in Rats with Non-alcoholic Fatty Liver Disease / 中国结合医学杂志

OBJECTIVE@#To evaluate the effect of Shilajit, a medicine of Ayurveda, on the serum changes in cytokines and adipokines caused by non-alcoholic fatty liver disease (NAFLD).@*METHODS@#After establishing fatty liver models by feeding a high-fat diet (HFD) for 12 weeks, 35 Wistar male rats were randomly divided into 5 groups, including control (standard diet), Veh (HFD + vehicle), high-dose Shilajit [H-Sh, HFD + 250 mg/(kg·d) Shilajit], low-dose Shilajit [L-Sh, HFD + 150 mg/(kg·d) Shilajit], and pioglitazone [HFD + 10 mg/(kg·d) pioglitazone] groups, 7 rats in each group. After 2-week of gavage administration, serum levels of glucose, insulin, interleukin 1beta (IL-1β), IL-6, IL-10, tumor necrosis factor-alpha (TNF-α), adiponectin, and resistin were measured, and insulin resistance index (HOMA-IR) was calculated.@*RESULTS@#After NAFLD induction, the serum level of IL-10 significantly increased and serum IL-1β, TNF-α levels significantly decreased by injection of both doses of Shilajit and pioglitazone (P<0.05). Increases in serum glucose level and homeostasis model of HOMA-IR were reduced by L-Sh and H-Sh treatment in NAFLD rats (P<0.05). Both doses of Shilajit increased adiponectin and decreased serum resistin levels (P<0.05).@*CONCLUSION@#The probable protective role of Shilajit in NAFLD model rats may be via modulating the serum levels of IL-1β, TNF-α, IL-10, adipokine and resistin, and reducing of HOMA-IR.

Aqueous date fruit efficiency as preventing traumatic brain deterioration and improving pathological parameters after traumatic brain injury in male rats

Objective: following traumatic brain injury, disruption of blood-brain-barrier and consequent brain edema are critical events which might lead to increasing intracranial pressure [ICP], and nerve damage. The current study assessed the effects of aqueous date fruit extract [ADFE] on the aforementioned parameters

Materials and Methods: in this experimental study, diffused traumatic brain injury [TBI] was generated in adult male rats using Marmarou's method. Experimental groups include two pre-treatment [oral ADFE, 4 and 8 mL/kg for 14 days], vehicle [distilled water, for 14 days] and sham groups. Brain edema and neuronal injury were measured 72 hours after TBI. Veterinary coma scale [VCS] and ICP were determined at -1, 4, 24, 48 and 72 hours after TBI. Differences among multiple groups were assessed using ANOVA. Turkey's test was employed for the ANOVA post-hoc analysis. The criterion of statistical significance was sign at P<0.05

Results: brain water content in ADFE-treated groups was decreased in comparison with the TBI+vehicle group. VCS at 24, 48 and 72 hours after TBI showed a significant increase in ADFE groups in comparison with the TBI+vehicle group. ICP at 24, 48 and 72 hours after TBI, was decreased in ADFE groups, compared to the TBI+vehicle. Brain edema, ICP and neuronal injury were also decreased in ADFE group, but VCS was increased following on TBI

Conclusion: ADFE pre-treatment demonstrated an efficient method for preventing traumatic brain deterioration and improving pathological parameters after TBI

Changes in neuromedin S and its receptor after traumatic brain injury in cycling rats

Animal studies indicate that gonadal steroids have prominent neuroprotective effects in several models of experimental traumatic brain injury (TBI). Neuromedin U (NMU) and neuromedin S (NMS) are regulatory peptides involved in inflammatory and stress responses, and modulation of the gonadotropic axis. Since steroid hormone levels change during the estrous cycle, we sought to determine whether variations in ovarian hormones would affect blood-brain barrier (BBB) permeability and brain levels of NMS, NMU, and neuromedin S receptor 2 in experimental TBI. Two groups (proestrus and nonproestrus) of female rats underwent diffuse TBI. At 24 hrs after TBI, results showed a significantly decrease in BBB permeability in traumatic-proestrus animals (TBI-P) in comparison to traumatic nonproestrus (TBI-NP) rats. Western blot analyzes demonstrated an enhanced expression of prepro-NMS in TBI-P compared with that in the TBI-NP group. Likewise, TBI-P rats exhibited significantly higher NMUR2 gene expression compared with those of TBI-NP, whereas no significant difference in brain NMU content was seen between sham and traumatic animals. Our findings indicate that diffuse TBI induces an increase in prepro-NMS and neuromedin S receptor 2 expression in traumatic-proestrus rats which may mediate the anti-edematous effects of gonadal hormones in proestrus rats following trauma.

Changes in neuromedin S and its receptor after traumatic brain injury in cycling rats

Animal studies indicate that gonadal steroids have prominent neuroprotective effects in several models of experimental traumatic brain injury (TBI). Neuromedin U (NMU) and neuromedin S (NMS) are regulatory peptides involved in inflammatory and stress responses, and modulation of the gonadotropic axis. Since steroid hormone levels change during the estrous cycle, we sought to determine whether variations in ovarian hormones would affect blood-brain barrier (BBB) permeability and brain levels of NMS, NMU, and neuromedin S receptor 2 in experimental TBI. Two groups (proestrus and nonproestrus) of female rats underwent diffuse TBI. At 24 hrs after TBI, results showed a significantly decrease in BBB permeability in traumatic-proestrus animals (TBI-P) in comparison to traumatic nonproestrus (TBI-NP) rats. Western blot analyzes demonstrated an enhanced expression of prepro-NMS in TBI-P compared with that in the TBI-NP group. Likewise, TBI-P rats exhibited significantly higher NMUR2 gene expression compared with those of TBI-NP, whereas no significant difference in brain NMU content was seen between sham and traumatic animals. Our findings indicate that diffuse TBI induces an increase in prepro-NMS and neuromedin S receptor 2 expression in traumatic-proestrus rats which may mediate the anti-edematous effects of gonadal hormones in proestrus rats following trauma.

effects of estrogen receptors' antagonist on brain edema, intracranial pressure and neurological outcomes after traumatic brain injury in rat

In previous studies, the neuroprotective effect of 17beta -estradiol in diffuse traumatic brain injury has been shown. This study used ICI 182,780, a non-selective estrogen receptor antagonist, to test the hypothesis that the neuroprotective effect of 17beta -estradiol in traumatic brain injury is mediated by the estrogen receptors. The ovariectomized rats were divided into eight groups. Brain injury was induced by Marmarou's method. Estrogen was injected 30 minutes after traumatic brain injury, and ICI 182,780 was injected before traumatic brain injury and also before estrogen treatment. In one group only ICI 182,780 was injected. The brain water content and Evans blue dye content were measured 24 and 5 hours after traumatic brain injury, respectively. The neurologic outcomes and intracranial pressure were assessed before, 4, and 24 hours after traumatic brain injury. Brain water content and Evans blue content were less in estrogen-treated group comparison to vehicle group. ICI 182,780 eliminated the effects of estrogen on brain edema and brain blood barrier permeability. Intracranial pressure was increased significantly after trauma, and estrogen decreased intracranial pressure at 4 and 24 hours after traumatic brain injury in comparison to vehicle. This inhibitory effect was also eliminated by treatment with ICI182,780. ICI 182,780 also inhibited the estrogen induced increase in neurologic outcomes following traumatic brain injury. However, the use of ICI 182,780 alone had no neuroprotective effect after traumatic brain injury. The results suggest that classical estrogen receptors have probably a role in the neuroprotective function of estrogen following traumatic brain injury

Traumatic brain injury has not prominent effects on cardiopulmonary indices of rat after 24 hours: hemodynamic, histopathology, and biochemical vidence

Accidents are the second reason for mortality and morbidity in Iran. Among them, brain injuries are the most important damage. Clarification of the effects of brain injuries on different body systems will help physicians to prioritize their treatment strategies. In this study, the effect of pure brain trauma on the cardiovascular system and lungs 24 hours post trauma was assessed. Male Wistar rats [n = 32] were divided into sham control and traumatic brain injury [TBI] groups. In TBI animals, under deep anesthesia, a blow to the head was induced by the fall of a 450 g weight from 2 m height. Twenty four hours later, heart electrocardiogram and functional indices, cardiac troponin I, IL-6, TNF-oc, IL-I[3 in tissue and serum, and the histopathology of heart and lung were assessed. The results showed that none of the functional, biochemical, inflammatory, and histopathology indices was statistically different between the two groups at 24 hours post TBI. Indices of impulse conduction velocity in atrium [P wave duration and P-R interval] were significantly longer in the TBI group. Overall, no important functional and histopathologic disturbances were found in heart and lung of TBI group after 24 hours. If the data is reproduced in human studies, the medical team could allocate their priority to treatment of brain disorders of the victim in the first 24 hours of pure TBI and postpone extensive assessment of heart and lung health indices to later time, thus reducing patient and health system expenditures

Neuroprotective antioxidant effect of sex steroid hormones in traumatic brain injury

The aim of the present study was to evaluate the effect of different doses of sex steroid hormones on brain edema, BBB permeability, brain antioxidant enzyme activity, and MDA level after traumatic brain injury [TBI] in ovarectomized [OVX] rats. Female rats were divided into six [One sham and 5 TBI] groups including: vehicle, estrogen in physiologic [33.3 microg/kg] and pharmacologic [1 mg/kg] doses, progesterone in physiologic [1.7 mg/kg] and pharmacological doses [8 mg/kg]. The results showed that compared to vehicle group, estrogen and progesterone groups showed significantly lower brain water content [P<0.001]. Evans blue content was significantly lower in both estrogen doses and in progesterone physiologic dose [P<0.001]. Evans blue content was significantly higher in progesterone pharmacologic dose [P<0.001]. Superoxide dismutase [SOD] activity was significantly higher in estrogen and progesterone pharmacologic doses [P<0.001]. Glutathione peroxidase [GPx] activity was significantly lower in estrogen physiologic dose [P<0.001]. It was concluded that the neuroprotective effect of different doses of sex steroid hormones after TBI, may be mediated by changes in oxidant agent activity

Chronic Opium Treatment Can Differentially Induce Brain and Liver Cells Apoptosis in Diabetic and Non-diabetic Male and Female Rats

It has been shown that some opium derivatives promote cell death via apoptosis. This study was designed to examine the influence of opium addiction on brain and liver cells apoptosis in male and female diabetic and non-diabetic Wistar rats. This experimental study was performed on normal, opium-addicted, diabetic and diabetic opium-addicted male and female rats. Apoptosis was evaluated by TUNEL and DNA fragmentation assays. Results of this study showed that apoptosis in opium-addicted and diabetic opium-addicted brain and liver cells were significantly higher than the both normal and diabetic rats. In addition, we found that apoptosis in brain cells of opium-addicted and diabetic opium-addicted male rats were significantly higher than opium-addicted and diabetic opium-addicted female, whereas apoptosis in liver cells of opium-addicted and diabetic opium-addicted female rats were significantly higher than opium-addicted and diabetic opium-addicted male. Overall, these results indicate that opium probably plays an important role in brain and liver cells apoptosis, therefore, leading neurotoxicity and hepatotoxicity. These findings also in away possibly means that male brain cells are more susceptible than female and interestingly liver of females are more sensitive than males in induction of apoptosis by opium.

effects of female sex steroids on gastric secretory responses of rat following traumatic brain injury

Gastric ulceration is induced by various forms of stress like surgery, ischemia and trauma. The female sex has more resistance to stress and the gastrointestinal lesions happen fewer than male sex. The purpose of this study was to evaluate the role of estradiol and progesterone on the gastric acid and pepsin levels following traumatic brain injury [TBI] induction. Materials and Methods Diffuse TBI was induced by Marmarou method in female rats. Rats randomly assigned into 9 groups: intact, OVX [ovarectomized rat], Sham+OVX, TBI [intact rats under TBI], TBI+OVX [ovarectomized rats under TBI] and treated OVX rats with vehicle [sesame oil], E2 [estradiol], P4 [progesterone] or E2+P4 combination. The acid content and pepsin levels of each gastric washout sample were measured 5 days after the TBI induction. There was no significant difference in gastric acid output between groups either after TBI induction or after treatment with E2 or P4 or E2+P4. Gastric pepsin levels were increased in Sham+OVX, TBI [P< 0.001] and TBI+OVX [P< 0.05] compared to intact group. Gastric pepsin levels were significantly lower in E2 and E2+ P4 treated rats than vehicle treated group [P< 0.01]. P4 treatment increased gastric pepsin level compared to TBI+OVX group [P< 0.05] and this increment was higher than rats that were treated with the E2 and E2+P4. These results suggest that protective effect of estradiol and E2+P4 combination against mucosal damage after TBI, might be mediated by inhibition of pepsin secretion

[Effects of different phases of estrous cycle on brain edema and neurological outcomes after severe traumatic brain injury in female rats]

Sex is one of the main recovery parameters after traumatic brain injury [TBI]. During estrus cycle phase, plasma estrogen and progesterone levels are very different. So in this study, we have assessed different effects of these phases in brain edema and neurological outcomes. Adult female Albino-N-MARI rats were divided into 6 groups [14 per group] including metestrus, diestrus, proestrus, estrus, ovarictomized [OVX] rats, and sham group [without TBI and ovary]. In all groups, brain water content for the measurement of brain edema, evans blue content for the measurement of brain vascular permeability, neurological scores, plasma estrogen and progesterone levels were assessed after severe TBI. Brain water content in diestrus, proestrus and estrus groups showed a significant decrease as compared with OVX group [P<0.001] but it was not significant in metestrus group. Evans blue content in proestrus group was lower than estrus and OVX groups [P<0.001]. Also, in diestrus group it was lower than OVX group [P<0.01].Neurological scores showed significant increases in proestrus group 4 hours after severe TBI as compared with metestrus, diestrus and OVX groups [P<0.01]. 24 hours after severe TBI, neurological scores in all groups were higher than OVX [P<0.001]. Brain water content and brain vascular permeability in sham group was lower than OVX, besides neurological scores in sham group were higher than OVX [P<0.001]. This study showed that brain edema, evans blue content and neurological scores after severe TBI are related to estrus cycle different phases. This could be related to differences in female sex hormones in different phases of estrus cycle

Thyroid function in mothers who gave birth to neonates with transient congenital hypothyroidism

To determine the thyroid status of mothers of newborns with primary congenital hypothyroidism. Thyroid function tests were carried out on 80 mothers of hypothyroid newborns and 80 mothers of non-hypothyroid newborns as control. The mean difference of the tests revealed that mothers of congenitally hypothyroid infants had a lower triiodothyronine resin uptake [T3RU] concentrations compared with the control population. The higher value of free thyroxin index [FTI] in case group showed a tendency to significance. The proportional frequency distribution showed; T3RU and triiodothyronine [T3] had a significant difference, and FTI showed a tendency to significance. There were no significant differences between; thyroid-stimulating hormone [TSH], thyroxine [T4] and anti-thyroid peroxidase antibodies [anti-TPO] in two groups. These results indicated that at least some cases of primary congenital hypothyroidism were attributable to the maternal thyroid disease. Therefore, we recommend that each pregnant woman should be assessed for thyroid function in region with a high prevalence of thyroid disease

Opium can differently alter blood glucose, sodium and potassium in male and female rats

To determine the effects of opium on serum glucose, potassium and sodium in male and female Wistar rat, opium solution [60 mg/kg] injected intraperitoneally and the same volume of distilled water was used as control [7 rats in each group]. Blood samples were collected at 0, 30, 60, 120, 240 and 360 minutes after injection from orbit cavity and the values of serum glucose, sodium [Na+] and potassium [K+] were measured. The data were then analyzed by the repeated measure ANOVA based on sex and case-control group. P < 0.05 considered as significant difference. Serum glucose increased significantly at 30, 60, 120 and 240 minutes after opium solution injection, in female rats compared to a control group. However, the male rats had this rise at 30, 60 and 120 minutes after opium solution injection compared to control group. While serum glucose in male rats was significantly higher than females at 30, 60 and 120 minutes, this value was higher in the female rats at 360 minutes. Therefore, serum glucose alterations following opium injection was significantly different in groups and in the sexes at different times. Sodium [Na+] rose at 60, 240 and 360 minutes significantly in all rats compared to control group. However, sodium alteration following opium injection was significantly different only between treated and control groups but sex-independent at all times. Potassium [K+] increased significantly at 60, 120, 240 and 360 minutes in male rats, compared to a control group. In female rats K+ significantly raised at 30, 120, 240 and 360 minutes. Therefore, the alteration of K+ in male and female rats was found time dependent and sex independent. According to our results, opium increased serum glucose in male and female rats differently, and it interferes with metabolic pathways differently on a gender dependent basis. Opium raised serum Na+ and K+, thus it interfere with water regulation and blood pressure via different mechanism

[ effect of sex steroid hormones on brain edema and intracranial pressure after experimental traumatic brain injury in rats]

We investigated the role of sex hormones on changes in brain edema intracranial pressure [ICP], cerebral perfusion pressure [CCP] after trauma brain injury [TBI] in ovarectomized female [OVX] rats. In this study female rats are divided into five groups. Control group [Intact] sham group and other groups include: vehicle, estrogen group [1mg/kg] and progesterone group [8 mg/kg] which on all groups TBI was induced by Marmarou method. 30 minutes after TBI, drugs were injected i.p. ICP was measured in spinal cord using a standard procedure. CPP was calculated by the mean arterial pressure [MAP] - ICP. Neurologic scores were measured by motor, eye and respiratory reflex. The results showed after TBI, water content was significantly lower in estrogen and progesterone groups [P<0.001] compared with vehicle group. Analysis showed a stable ICP up to 24 hours. The ICP in estrogen and progesterone groups was significantly decreased at 4 and 24 hours as compared to vehicle group [P<0.001in both cases]. The CPP at 24 hours after TBI, significantly increased in estrogen and progesterone groups compared with vehicle [P<0.001]. Also after TBI, neurologic scores was significantly higher in estrogen and progesterone groups as compared with vehicle [at 1 hours P<0.05, and at 24hours P<0.001 for estrogen], [at 1 hours P<0.01 for progesterone]. Our findings indicated an improvement of ICP, CPP and neurologic scores produced by pharmacologic doses of estrogen and progesterone after TBI in OVX rat. These effects may be contribute to neuroprotective effects of these hormones

W-7 [a calmodulin antagonist] inhibits carrageenan-induced paw edema in intact and adrenalectomized rats

The aim of present study was to investigate the effect of W-7, a calmodulin antagonist, on a carrageenan-induced rat's paw edema. W-7[50 micro Mol/kg] was given intraperitoneally synchronous with the intraplantar injection of 0.1 ml of 0.5% carrageenan solution. After for hours, paw edema was assessed by calculating the paw volume changes which measured by hydroplethysmometer. In adrenalectomized rats, both adrenal glands were removed. Treatment of animals with W-7 reduced carrageenan-induced paw edema by 50%. In adrenalectomized rats, W-7 was also effective in reduction of paw edema [52%]. There was no significant difference between the effect of W-7 in adrenalectomized and control animals. Our findings suggest that W-7 as calmodulin inhibitor reduce carrageenan-induced paw edema and the inhibitory effect W-7 on edema formation appears not to be dependent on adrenal function

Survey of changes in complete blood count and red cell indices of whole blood incubated in vitro at different temperatures up to 48 hours

Complete blood count [CBC] is one of the most common and conventional blood test that physicians usually request. However the results of this test are affected by different factors such as, the temperature and duration of incubation, therefore the aim of this survey was to evaluate the effect of temperature and time of incubation on CBC, red blood cells [RBC] indices and white blood cells [WBC] differential count. In a cross-sectional study, blood samples were taken from 30 healthy medical students of Rafsanjan University [15 males and 15 females]. The samples divided into three parts; CBC were done on the samples up to 48 hours incubation at temperature of 25, 30, and 37°C at the time of sampling, and after 2, 8, 24 and 48 hours. Data were statistically analyzed and the following results were obtained. RBC count, hematocrit, MCH, percent of monocytes and eosinophils were constant in different temperatures, WBC count, MCHC, hemoglobin, platelets count, the percent of lymphocytes and neutrophils were constant up to 24 hours and then tend to increase with increasing temperature except lymphocytes percent that tend to decrease. MCV decreased with increasing temperature up to 8 hours and then significantly increased [from 83.89 to 87.50 fmol/1, p<0.00l]. WBC, hematocrit, MCV, platelets count, and neutrophils' percent tend to increase by the time of incubation, but RBC count, MCHC, lymphocytes' percent decreased. Hemoglobin, MCH, and the percent of monocytes and eosinophils were constant. The finding of this survey showed that some of CBC parameters can be changed with the incubation, therefore it is better to do the CBC test after blood taking as soon as possible

Inhibition of proteinase 3 [PR3] by suramin and fetal calf serum [FCS]: effect of PR3 and suramin on Chinese hamster ovary cells [CHO-Cells]

Proteinase 3 [PR3] is a lysosomal protease that is stored in azurophilic granules neutrophilic granulocytes and monocytes. A number of inhibitors for this proteinase are reported. Comprehensive studies on the inhibitory effect of suramin and heat treated fetal calf serum [pound GFCS] on PR3 have not been reported. It has been reported that PR3 is able to destroy the cytoskeletal integral proteins, but we have not find any reports which showed the effect of this protease on Chinese hamster ovary cells [CHO-cells] in culture medium. Suramin has proven to be useful as an antitumor drug, but there was not any report on the effect of suramin on CHO-cells. The effects of various concentrations of pound GFCS [from 0.5% up to 10%] and suramin [from 0.8 pound gM up to 100 pound gM] on PR3 and different concentrations of suramin [from 0.8 pound gM up to 1000 pound gM] on CHO-cells were investigated. Data analysis were performed by, Kolmogorov-Smirnov test, ANOVA test and Tukey HSD post tests. Results showed that pound GFCS and suramin have an inhibitory effect on PR3 and these effects increased with increasing the concentration significantly [p < 0.01]. PR3 with the concentration of 2.2 Unit/ml has no effect on CHO-cells. Although suramin with the concentration of less than 125 poundgM cell growth retarded for only a few hours, but with the concentration of 125 to 250 pound gM inhibit the cell growth for a week, and after that cells gain normal growth gradually. Suramin with concentration of more than 500 pound gM inhibited the cell growth completely. Although suramin reversibly inhibit the PR3 activity but in concentration of less than 250 pound gM it had no long-term effect on CHO-cells. Therefore it can be used in the investigation of proteases. There were unknown components in pound GFCS, which cause the inhibition of PR3 activity. This finding is very important in PR3 production in culture medium. However CHO-cells are resistant to PR3 and suramin in low concentration

Transfection of 6-23 a rat thyroid carcinoma cell line, with the neuropeptide Y cDNA

Neuropeptide Y [NPY] is a 36 amino acid peptide found throughout the central and peripheral nervous system of rat and human. NPY has been proposed to play an important role in satiety. The aim of this study was to produce cell lines that secrete high levels of bioactive NPY. For this purpose, the complementary DNA [cDNA] that encodes NPY was isolated by PCR. The cDNA was then cloned into pCEP4, to form pCEP4NPY. 6-23 cells were transfected with pCEP4NPY by electroporation. Transfected cells were selected by the addition of hygromycin B to the culture medium. Resistant colonies were picked and transferred to 96-well plates. The medium was tested for IR-NPY using a specific NPY radioimmunoassay [RIA]. The IR-NPY secreted by the cells was characterized by sephadex G50 chromatography and reversed phase fast protein liquid chromatography [FPLC]. It was found to co-elute with the synthetic standard in both cases. RNA was extracted from the cells and subjected to Northern blot analysis using labeled NPY cDNA as a probe. The cells were found to express high levels of NPY at mRNA levels